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In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model

  • Authors:
    • Takashi Nishizawa
    • Kiyotaka Nakano
    • Etsuko Fujii
    • Daisuke Komura
    • Yoshie Kuroiwa
    • Chisako Ishimaru
    • Makoto Monnai
    • Hiroyuki Aburatani
    • Shumpei Ishikawa
    • Masami Suzuki
  • View Affiliations / Copyright

    Affiliations: Department of Research, Forerunner Pharma Research Co., Ltd., Komaba Open Laboratory, The University of Tokyo, Tokyo 153‑8904, Japan, Department of Genomic Pathology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo 113‑8510, Japan, Chugai Research Institute for Medical Science Co., Ltd., Kamakura, Kanagawa 247‑8530, Japan, Genome Science Division, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo 153‑8904, Japan
    Copyright: © Nishizawa et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Pages: 1745-1754
    |
    Published online on: September 3, 2019
       https://doi.org/10.3892/or.2019.7300
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Abstract

Ras homolog family member A (RHOA) mutations are driver genes in diffuse‑type gastric cancers (DGCs), and we previously revealed that RHOA mutations contribute to cancer cell survival and cell migration through their dominant negative effect on Rho‑associated kinase (ROCK) signaling in vitro. However, how RHOA mutations contribute to DGC development in vivo is poorly understood. In the present study, the contribution of RHOA mutations to tumor morphology was investigated using an orthotopic xenograft model using the gastric cancer cell line MKN74, in which wild‑type (WT) or mutated (Y42C and Y42S) RHOA had been introduced. When we conducted RNA sequencing to distinguish between the genes expressed in human tumor tissues from those in mouse stroma, the expression profiles of the tumors were clearly divided into a Y42C/Y42S group and a mock/WT group. Through gene set enrichment analysis, it was revealed that inflammation‑ and hypoxia‑related pathways were enriched in the mock/WT tumors; however, cell metabolism‑ and cell cycle‑related pathways such as Myc, E2F, oxidative phosphorylation and G2M checkpoint were enriched in the Y42C/Y42S tumors. In addition, the gene set related to ROCK signaling inhibition was enriched in the RHOA‑mutated group, which indicated that a series of events are related to ROCK inhibition induced by RHOA mutations. Histopathological analysis revealed that small tumor nests were more frequent in RHOA mutants than in the mock or WT group. In addition, increased blood vessel formation and infiltration of macrophages within the tumor mass were observed in the RHOA mutants. Furthermore, unlike mock/WT, the RHOA‑mutated tumor cells had little antitumor host reaction in the invasive front, which is similar to the pattern of mucosal invasion in clinical RHOA‑mutated DGC. These transcriptome and pathological analyses revealed that mutated RHOA functionally contributes to the acquisition of DGC features, which will accelerate our understanding of the contribution of RHOA mutations in DGC biology and the development of further therapeutic strategies.
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Copy and paste a formatted citation
Spandidos Publications style
Nishizawa T, Nakano K, Fujii E, Komura D, Kuroiwa Y, Ishimaru C, Monnai M, Aburatani H, Ishikawa S, Suzuki M, Suzuki M, et al: In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model. Oncol Rep 42: 1745-1754, 2019.
APA
Nishizawa, T., Nakano, K., Fujii, E., Komura, D., Kuroiwa, Y., Ishimaru, C. ... Suzuki, M. (2019). In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model. Oncology Reports, 42, 1745-1754. https://doi.org/10.3892/or.2019.7300
MLA
Nishizawa, T., Nakano, K., Fujii, E., Komura, D., Kuroiwa, Y., Ishimaru, C., Monnai, M., Aburatani, H., Ishikawa, S., Suzuki, M."In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model". Oncology Reports 42.5 (2019): 1745-1754.
Chicago
Nishizawa, T., Nakano, K., Fujii, E., Komura, D., Kuroiwa, Y., Ishimaru, C., Monnai, M., Aburatani, H., Ishikawa, S., Suzuki, M."In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model". Oncology Reports 42, no. 5 (2019): 1745-1754. https://doi.org/10.3892/or.2019.7300
Copy and paste a formatted citation
x
Spandidos Publications style
Nishizawa T, Nakano K, Fujii E, Komura D, Kuroiwa Y, Ishimaru C, Monnai M, Aburatani H, Ishikawa S, Suzuki M, Suzuki M, et al: In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model. Oncol Rep 42: 1745-1754, 2019.
APA
Nishizawa, T., Nakano, K., Fujii, E., Komura, D., Kuroiwa, Y., Ishimaru, C. ... Suzuki, M. (2019). In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model. Oncology Reports, 42, 1745-1754. https://doi.org/10.3892/or.2019.7300
MLA
Nishizawa, T., Nakano, K., Fujii, E., Komura, D., Kuroiwa, Y., Ishimaru, C., Monnai, M., Aburatani, H., Ishikawa, S., Suzuki, M."In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model". Oncology Reports 42.5 (2019): 1745-1754.
Chicago
Nishizawa, T., Nakano, K., Fujii, E., Komura, D., Kuroiwa, Y., Ishimaru, C., Monnai, M., Aburatani, H., Ishikawa, S., Suzuki, M."In vivo effects of mutant RHOA on tumor formation in an orthotopic inoculation model". Oncology Reports 42, no. 5 (2019): 1745-1754. https://doi.org/10.3892/or.2019.7300
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